Skip to main content
SpringerLink
Log in

Natural killer cell is a major producer of interferon γ that is critical for the IL-12-induced anti-tumor effect in mice

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Although the anti-tumor effect of IL-12 is mediated mostly by IFNγ, which cell types most efficiently produce IFNγ and therefore initiate or promote the anti-tumor effect of IL-12 has not been clearly determined. In the present study, we demonstrated hydrodynamic injection of the IL-12 gene led to prolonged IFNγ production, NK-cell activation and complete inhibition of liver metastasis of CT-26 colon cancer cells in wild-type mice, but not in IFNγ knockout mice. NK cells expressed higher levels of STAT4 and upon IL-12 administration displayed stronger STAT4 phosphorylation and IFNγ production than non-NK cells. Adoptive transfer of wild-type NK cells into IFNγ knockout mice restored IL-12-induced IFNγ production, NK-cell activation and anti-tumor effect, whereas transfer of the same number of wild-type non-NK cells did not. In conclusion, NK cells are predominant producers of IFNγ that is critical for IL-12 anti-tumor therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Kobayashi M, Fitz L, Ryan M, Hewick RM, Clark SC, Chan S, Loudon R, Sherman F, Perussia B, Trinchieri G (1989) Identification and purification of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic effects on human lymphocytes. J Exp Med 170(3):827–845

    Article  CAS  PubMed  Google Scholar 

  2. Stern AS, Podlaski FJ, Hulmes JD, Pan YC, Quinn PM, Wolitzky AG, Familletti PC, Stremlo DL, Truitt T, Chizzonite R, Gately MK (1990) Purification to homogeneity and partial characterization of cytotoxic lymphocyte maturation factor from human B-lymphoblastoid cells. Proc Natl Acad Sci USA 87(17):6808–6812

    Article  CAS  PubMed  Google Scholar 

  3. Watford WT, Moriguchi M, Morinobu A, O’Shea JJ (2003) The biology of IL-12: coordinating innate and adaptive immune responses. Cytokine Growth Factor Rev 14(5):361–368

    Article  CAS  PubMed  Google Scholar 

  4. Trinchieri G (2003) Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat Rev Immunol 3(2):133–146

    Article  CAS  PubMed  Google Scholar 

  5. Colombo MP, Trinchieri G (2002) Interleukin-12 in anti-tumor immunity and immunotherapy. Cytokine Growth Factor Rev 13(2):155–168

    Article  CAS  PubMed  Google Scholar 

  6. Del Vecchio M, Bajetta E, Canova S, Lotze MT, Wesa A, Parmiani G, Anichini A (2007) Interleukin-12: biological properties and clinical application. Clin Cancer Res 13(16):4677–4685

    Article  CAS  PubMed  Google Scholar 

  7. Wigginton JM, Gruys E, Geiselhart L, Subleski J, Komschlies KL, Park JW, Wiltrout TA, Nagashima K, Back TC, Wiltrout RH (2001) IFN-gamma and Fas/FasL are required for the antitumor and antiangiogenic effects of IL-12/pulse IL-2 therapy. J Clin Invest 108(1):51–62

    CAS  PubMed  Google Scholar 

  8. Lee JC, Kim DC, Gee MS, Saunders HM, Sehgal CM, Feldman MD, Ross SR, Lee WM (2002) Interleukin-12 inhibits angiogenesis and growth of transplanted but not in situ mouse mammary tumor virus-induced mammary carcinomas. Cancer Res 62(3):747–755

    CAS  PubMed  Google Scholar 

  9. Brunda MJ, Luistro L, Warrier RR, Wright RB, Hubbard BR, Murphy M, Wolf SF, Gately MK (1993) Antitumor and antimetastatic activity of interleukin 12 against murine tumors. J Exp Med 178(4):1223–1230

    Article  CAS  PubMed  Google Scholar 

  10. Cui J, Shin T, Kawano T, Sato H, Kondo E, Toura I, Kaneko Y, Koseki H, Kanno M, Taniguchi M (1997) Requirement for Valpha14 NKT cells in IL-12-mediated rejection of tumors. Science 278(5343):1623–1626

    Article  CAS  PubMed  Google Scholar 

  11. Zilocchi C, Stoppacciaro A, Chiodoni C, Parenza M, Terrazzini N, Colombo MP (1998) Interferon gamma-independent rejection of interleukin 12-transduced carcinoma cells requires CD4 + T cells and Granulocyte/Macrophage colony-stimulating factor. J Exp Med 188(1):133–143

    Article  CAS  PubMed  Google Scholar 

  12. Kodama T, Takeda K, Shimozato O, Hayakawa Y, Atsuta M, Kobayashi K, Ito M, Yagita H, Okumura K (1999) Perforin-dependent NK cell cytotoxicity is sufficient for anti-metastatic effect of IL-12. Eur J Immunol 29(4):1390–1396

    Article  CAS  PubMed  Google Scholar 

  13. Takeda K, Hayakawa Y, Atsuta M, Hong S, Van Kaer L, Kobayashi K, Ito M, Yagita H, Okumura K (2000) Relative contribution of NK and NKT cells to the anti-metastatic activities of IL-12. Int Immunol 12(6):909–914

    Article  CAS  PubMed  Google Scholar 

  14. Ogawa M, Yu WG, Umehara K, Iwasaki M, Wijesuriya R, Tsujimura T, Kubo T, Fujiwara H, Hamaoka T (1998) Multiple roles of interferon-gamma in the mediation of interleukin 12-induced tumor regression. Cancer Res 58(11):2426–2432

    CAS  PubMed  Google Scholar 

  15. Subleski JJ, Hall VL, Back TC, Ortaldo JR, Wiltrout RH (2006) Enhanced antitumor response by divergent modulation of natural killer and natural killer T cells in the liver. Cancer Res 66(22):11005–11012

    Article  CAS  PubMed  Google Scholar 

  16. Kubin M, Kamoun M, Trinchieri G (1994) Interleukin 12 synergizes with B7/CD28 interaction in inducing efficient proliferation and cytokine production of human T cells. J Exp Med 180(1):211–222

    Article  CAS  PubMed  Google Scholar 

  17. Yoshimoto T, Okamura H, Tagawa YI, Iwakura Y, Nakanishi K (1997) Interleukin 18 together with interleukin 12 inhibits IgE production by induction of interferon-gamma production from activated B cells. Proc Natl Acad Sci USA 94(8):3948–3953

    Article  CAS  PubMed  Google Scholar 

  18. Lauwerys BR, Renauld JC, Houssiau FA (1999) Synergistic proliferation and activation of natural killer cells by interleukin 12 and interleukin 18. Cytokine 11(11):822–830

    Article  CAS  PubMed  Google Scholar 

  19. Takehara T, Uemura A, Tatsumi T, Suzuki T, Kimura R, Shiotani A, Ohkawa K, Kanto T, Hiramatsu N, Hayashi N (2007) Natural killer cell-mediated ablation of metastatic liver tumors by hydrodynamic injection of IFNalpha gene to mice. Int J Cancer 120(6):1252–1260

    Article  CAS  PubMed  Google Scholar 

  20. Watanabe M, Fenton RG, Wigginton JM, McCormick KL, Volker KM, Fogler WE, Roessler PG, Wiltrout RH (1999) Intradermal delivery of IL-12 naked DNA induces systemic NK cell activation and Th1 response in vivo that is independent of endogenous IL-12 production. J Immunol 163(4):1943–1950

    CAS  PubMed  Google Scholar 

  21. Takehara T, Suzuki T, Ohkawa K, Hosui A, Jinushi M, Miyagi T, Tatsumi T, Kanazawa Y, Hayashi N (2006) Viral covalently closed circular DNA in a non-transgenic mouse model for chronic hepatitis B virus replication. J Hepatol 44(2):267–274

    Article  CAS  PubMed  Google Scholar 

  22. Shin T, Nakayama T, Akutsu Y, Motohashi S, Shibata Y, Harada M, Kamada N, Shimizu C, Shimizu E, Saito T, Ochiai T, Taniguchi M (2001) Inhibition of tumor metastasis by adoptive transfer of IL-12-activated Valpha14 NKT cells. Int J Cancer 91(4):523–528

    Article  CAS  PubMed  Google Scholar 

  23. Thierfelder WE, van Deursen JM, Yamamoto K, Tripp RA, Sarawar SR, Carson RT, Sangster MY, Vignali DA, Doherty PC, Grosveld GC, Ihle JN (1996) Requirement for Stat4 in interleukin-12-mediated responses of natural killer and T cells. Nature 382(6587):171–174

    Article  CAS  PubMed  Google Scholar 

  24. Kaplan MH, Sun YL, Hoey T, Grusby MJ (1996) Impaired IL-12 responses and enhanced development of Th2 cells in Stat4-deficient mice. Nature 382(6587):174–177

    Article  CAS  PubMed  Google Scholar 

  25. Morinobu A, Gadina M, Strober W, Visconti R, Fornace A, Montagna C, Feldman GM, Nishikomori R, O’Shea JJ (2002) STAT4 serine phosphorylation is critical for IL-12-induced IFN-gamma production but not for cell proliferation. Proc Natl Acad Sci USA 99(19):12281–12286

    Article  CAS  PubMed  Google Scholar 

  26. Comes A, Di Carlo E, Musiani P, Rosso O, Meazza R, Chiodoni C, Colombo MP, Ferrini S (2002) IFN-gamma-independent synergistic effects of IL-12 and IL-15 induce anti-tumor immune responses in syngeneic mice. Eur J Immunol 32(7):1914–1923

    Article  CAS  PubMed  Google Scholar 

  27. Hafner M, Falk W, Echtenacher B, Mannel DN (1999) Interleukin-12 activates NK cells for IFN-gamma-dependent and NKT cells for IFN-gamma-independent antimetastatic activity. Eur Cytokine Netw 10(4):541–548

    CAS  PubMed  Google Scholar 

  28. Komita H, Homma S, Saotome H, Zeniya M, Ohno T, Toda G (2006) Interferon-gamma produced by interleukin-12-activated tumor infiltrating CD8 + T cells directly induces apoptosis of mouse hepatocellular carcinoma. J Hepatol 45(5):662–672

    Article  CAS  PubMed  Google Scholar 

  29. Segal JG, Lee NC, Tsung YL, Norton JA, Tsung K (2002) The role of IFN-gamma in rejection of established tumors by IL-12: source of production and target. Cancer Res 62(16):4696–4703

    CAS  PubMed  Google Scholar 

  30. Nastala CL, Edington HD, McKinney TG, Tahara H, Nalesnik MA, Brunda MJ, Gately MK, Wolf SF, Schreiber RD, Storkus WJ, Lotze MT (1994) Recombinant IL-12 administration induces tumor regression in association with IFN-gamma production. J Immunol 153(4):1697–1706

    CAS  PubMed  Google Scholar 

  31. Doherty DG, O’Farrelly C (2000) Innate and adaptive lymphoid cells in the human liver. Immunol Rev 174:5–20

    Article  CAS  PubMed  Google Scholar 

  32. Seki S, Habu Y, Kawamura T, Takeda K, Dobashi H, Ohkawa T, Hiraide H (2000) The liver as a crucial organ in the first line of host defense: the roles of Kupffer cells, natural killer (NK) cells and NK1.1 Ag + T cells in T helper 1 immune responses. Immunol Rev 174:35–46

    Article  CAS  PubMed  Google Scholar 

  33. Car BD, Eng VM, Lipman JM, Anderson TD (1999) The toxicology of interleukin-12: a review. Toxicol Pathol 27(1):58–63

    Article  CAS  PubMed  Google Scholar 

  34. Biber JL, Jabbour S, Parihar R, Dierksheide J, Hu Y, Baumann H, Bouchard P, Caligiuri MA, Carson W (2002) Administration of two macrophage-derived interferon-gamma-inducing factors (IL-12 and IL-15) induces a lethal systemic inflammatory response in mice that is dependent on natural killer cells but does not require interferon-gamma. Cell Immunol 216(1–2):31–42

    Article  CAS  PubMed  Google Scholar 

  35. Tsushima H, Ito N, Tamura S, Matsuda Y, Inada M, Yabuuchi I, Imai Y, Nagashima R, Misawa H, Takeda H, Matsuzawa Y, Kawata S (2001) Circulating transforming growth factor beta 1 as a predictor of liver metastasis after resection in colorectal cancer. Clin Cancer Res 7(5):1258–1262

    CAS  PubMed  Google Scholar 

  36. Okumoto K, Hattori E, Tamura K, Kiso S, Watanabe H, Saito K, Saito T, Togashi H, Kawata S (2004) Possible contribution of circulating transforming growth factor-beta1 to immunity and prognosis in unresectable hepatocellular carcinoma. Liver Int 24(1):21–28

    Article  CAS  PubMed  Google Scholar 

  37. Chau GY, Wu CW, Lui WY, Chang TJ, Kao HL, Wu LH, King KL, Loong CC, Hsia CY, Chi CW (2000) Serum interleukin-10 but not interleukin-6 is related to clinical outcome in patients with resectable hepatocellular carcinoma. Ann Surg 231(4):552–558

    Article  CAS  PubMed  Google Scholar 

  38. Galizia G, Lieto E, De Vita F, Romano C, Orditura M, Castellano P, Imperatore V, Infusino S, Catalano G, Pignatelli C (2002) Circulating levels of interleukin-10 and interleukin-6 in gastric and colon cancer patients before and after surgery: relationship with radicality and outcome. J Interferon Cytokine Res 22(4):473–482

    Article  CAS  PubMed  Google Scholar 

  39. Meadows SK, Eriksson M, Barber A, Sentman CL (2006) Human NK cell IFN-gamma production is regulated by endogenous TGF-beta. Int Immunopharmacol 6(6):1020–1028

    Article  CAS  PubMed  Google Scholar 

  40. Caruso M, Pham-Nguyen K, Kwong YL, Xu B, Kosai KI, Finegold M, Woo SL, Chen SH (1996) Adenovirus-mediated interleukin-12 gene therapy for metastatic colon carcinoma. Proc Natl Acad Sci USA 93(21):11302–11306

    Article  CAS  PubMed  Google Scholar 

  41. Barajas M, Mazzolini G, Genove G, Bilbao R, Narvaiza I, Schmitz V, Sangro B, Melero I, Qian C, Prieto J (2001) Gene therapy of orthotopic hepatocellular carcinoma in rats using adenovirus coding for interleukin 12. Hepatology 33(1):52–61

    Article  CAS  PubMed  Google Scholar 

  42. Sangro B, Mazzolini G, Ruiz J, Herraiz M, Quiroga J, Herrero I, Benito A, Larrache J, Pueyo J, Subtil JC, Olague C, Sola J et al (2004) Phase I trial of intratumoral injection of an adenovirus encoding interleukin-12 for advanced digestive tumors. J Clin Oncol 22(8):1389–1397

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Dr. Morihiro Watanabe (Laboratory of Experimental Immunology, Division of Basic Sciences, National Cancer Institute-Frederick Cancer Research and Development Center) for providing the pCMV-IL-12 plasmid, Dr. Yoichiro Iwakura (University of Tokyo, Institute of Medical Science) for providing GKO mice.

Author information

Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Akio Uemura, Tetsuo Takehara, Takuya Miyagi, Takahiro Suzuki, Tomohide Tatsumi, Kazuyoshi Ohkawa, Tatsuya Kanto, Naoki Hiramatsu & Norio Hayashi

Authors
  1. Akio Uemura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tetsuo Takehara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takuya Miyagi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takahiro Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tomohide Tatsumi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kazuyoshi Ohkawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tatsuya Kanto
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Naoki Hiramatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Norio Hayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Norio Hayashi.

Additional information

A. Uemura and T. Takehara contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (TIFF 236 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Uemura, A., Takehara, T., Miyagi, T. et al. Natural killer cell is a major producer of interferon γ that is critical for the IL-12-induced anti-tumor effect in mice. Cancer Immunol Immunother 59, 453–463 (2010). https://doi.org/10.1007/s00262-009-0764-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-009-0764-x

Keywords

Search

Navigation